BACKGROUND: Chinese herbs of common yam rhizome, balsampear fruit and bagasse fiber have good effects on decreasing blood glucose and lipid, but its mechanism is unclear.OBJECTIVE: To observe the effect of compound preparation of common yam rhizome and balsampear fruit on blood glucose, lipid, blood insulin and anti-infection of rats with type 2 diabetes mellitus (DM).DESIGN: Randomized controlled animal study.SETTING: Technological Developing Center, Pharmacological Department, Experimental Animal Center, and Central Laboratory of Guangdong Medical College.MATERIALS: A total of 80 female SD rats with 4 months old and of SPF grade were selected in this study. Flumamine (Jilin Dongbeiya Pharmaceutical Co., Ltd., batch number: 040126); total cholesterol (TC), high density lipoprotein cholesterol (HDL-C) and triacylglycerol (TG) (Beijing Zhongsheng Biotechnological Co. Ltd.); Surestep Life scan blood glucose meter and test paper (Johnson Company, USA); insulin radio-immunity kit (Shanghai Navy Medical Institute); UV-3010 ultraviolet spectrophotometer (Japan Shimadzu); compound preparation of common yam rhizome and balsampear fruit (Technological Developing Center of Guangdong Medical College, batch number: 040321); suspension was extracted from common yam rhizome, balsampear fruit and dietary fiber of bagasse through water with 1 kg/L raw materials.METHODS: Animal breeding and samples collecting were carried out in the Experimental Animal Center and Technological Developing Central Laboratory of Guangdong Medical College from June 2004 to December 2005; meanwhile, detection of marker was carried out in the Pharmacological Department and Central Laboratory of Guangdong Medical College. ①Twenty rats were randomly selected as normal control group and perfused with 5 mL/kg saline every day. Other 60 rats were perfused with 5 mL/kg fat emulsion once a day for 4 weeks, and then, rats were fasted for 12hours and peritoneally injected with 2 g/L streptozotocin (30 mg/kg). Rats in normal control group were peritoneally injected with the same volume of citromalic acid buffer. Three days later, blood glucose was measured ranomly and measured again after 2 weeks. If level of blood glucose was igher than 13.5 mmol/L or urinal glucose was ＞ ++ for two weeks, the models were successful (n=48). ② According to random lot method, 48 rats were divided into three groups: model group, flumamine group and comound preparation group with 16 in each group. Rats in model group were perfused with 5 mL/kg fat emulsion; moreover, rats in flumamine groupr and compound preparation group were perfused with 5 mL/kg fat emulsion and then with 1 mg/kg flumamine and 5 mL/kg compound preparation including 1 kg/L raw drug, respectively. Rats in normal control group were perfused with 5 mL/kg saline. All rats in each group were perfused once a day for 6 weeks in total. ③ Value of blood glucose was measured at one day before the experiment finished. Twelve hours after fasting, level of plasma insulin was measured with radio-immunity method; levels of plasma total protein and albumin were measured with spectrophotography; levels of TG, TC and HDL-C were measured with the related kits. ④ Measurement data were compared with analysis of variance (ANOVA). Levene's test was firstly used to evaluate regularity of variance. Bonferroni test was used for regular variance; however, Tamhane's T2 was used.MAIN OUTCOME MEASURES: Effect of compound preparation on levels of blood glucose, insulin, lipid and plasma protein of rats with type 2 DM.RESULTS: Twelve rats were lost because of failure in modeling, and 4rats in model group and 2 in flumamine group died during the experiment,respectively. Therefore, 62 rats were involved in the final analysis. ①Measurement of fasting blood glucose and plasma insulin: Value of fasting blood glucose in normal control group was lower than that in other three groups (t=2.673-4.224, P ＜ 0.05-0.01), but level of plasma insulin was higher than that in other three groups (t=3.780-5.824, P ＜ 0.05-0.01).Fasting insulin in model group was lower than that in compound prepara tion group (t=2.825, P ＜ 0.05); fasting blood glucose was higher than that in flumamine group and compound preparation group (t=3.906, 3.056, P ＜ 0.05); * level of insulin in flumamine group was lower than that in compound preparation group (t=3.014, P ＜ 0.05); level of fasting blood glucose in flumamine group was close to that in compound preparation group (P ＞ 0.05). ② Measurement of lipid: Levels of TC and TG in normal control group were lower than those in other three groups, but level of HDL-C was higher than that in other three groups (t=2.521-4.892, P ＜ 0.05-0.01).Plasma TC in model group was higher than that in flumamine group and compound preparation group (t=2.466-2.512, P ＜ 0.05), value of TG was higher than that in compound preparation group (t=2.612, P ＜ 0.05), and level of HDL-C was lower than that in compound preparation group (t =3.688, P ＜ 0.05). Plasma TG in flumamine group was higher than that in compound preparation group (t=2.620, P ＜ 0.05). ③ Measurement of plasma protein: Levels of plasma total protein were close to each other (P ＞ 0.05). Plasma albumin in normal control group was higher than that in model group and flumamine group (t=3.773, 3.104, P ＜ 0.05), but that was close to that in compound preparation group (P ＞ 0.05). Ratio between albumin and globulin in normal oln that in other groups (t=2.830-3.056, P ＜ 0.05). Level of plasma albumin and ratio between albumin and globulin were lower in model group than those in compound preparation group (t=2.604, 3.808, P ＜ 0.05).CONCLUSION: Compound preparation can decrease levels of blood glucose and lipid, increase content of insulin, and improve anti-infection ability of rats with type 2 DM.

Biodiesel, a nontoxic,cleaning, renewable and biodegradable fuel, is expected as a substitute for conventional fossil diesel. There are three main approaches to produce biodiesel, alkali-catalysis processing, enzymatic-catalysis processing and supercritical processing. With the unique property of energy-saving and environment-friendly, enzymatic-catalysis appears a great potential for industrial application. The main bottleneck of this technology is high cost and low stability of the lipase, as well as the inactivation of lipase by methanol and so on. To settle the problem, several methods have been used including the fixed-bed bioreactor, enzyme immobilized processing, whole-cell biocatalyst, changing addition method of methanol, developing of novel acyl acceptor, enhancing methanol resistance of lipase. The main problems and the relative strategy research of the enzymatic-catalysis technology were sum up.

Objective To assess the value of salivary gland scintigraphy in diagnosis of Sj(o)gren's syndrome (SS).Methods A total of 44 patients with clinically suspicious SS were included.The data of salivary gland scintigraphy were retrospectively analyzed and the time-radioactivity curve (TAC) was obtained by outlining ROI in bilateral parotid glands and submaxillary glands.Uptake index (UI) and excretion fraction (EF) were defined.Both UI and EF were compared with the visual assessment and final diagnosis respectively.Results UI and EF of bilateral parotid glands and submaxillary glands in SS patients were significantly lower than those in non-SS patients (all P＜0.05).The impaired salivary gland function was classified as 0-3 grades by visual assessment.The UI of bilateral parotid glands and submaxillary glands were negatively correlated with the qualitative classification.While there were no significant correlations between EF and qualitative classification (all P＞0.05),except for that of right submaxillary gland (r=-0.312,P=0.039).The comprehensive diagnostic efficacy of UI on SS patients was higher than those of visual assessment,but their area under curves of ROC were not significantly different (all P＞0.05).Conclusion UI and EF can effectively evaluate salivary gland function and serve as objective tools to distinguish patients with SS.

Objective To evaluate the outcome of combination of intensive preconditioning regimen allo-HSCT with imatinib for treatment of Ph chromosome positive acute lymphocyte leukemia (ALL). Methods Between 2009 and 2010, 8 patients diagnosed as Ph+ ALL received allo-HSCT from HLA identical sibling during complete remission. Imatinib was added into the therapies of 5 patients.Seven patients received the intensive preconditioning regimen based on BuCy2, one patient received the regimen of TBI-Cy. A median of 6. 02 × 108/kg mononuclear cells and 3. 14 × 106/kg CD34+ cells were transfused. GVHD prophylaxis included cyclosporine A and methotrexate. Results All patients were well tolerant to the regimen without serious regimen-related toxicity. The median time of ANC≥0. 5 × 109/L was 15. 5 days, and that of PLT≥20 × 109/L was 19 days. Thirty days after allo-HSCT, all patients got donor engraftment successfully. Among 8 cases, 4 cases presented acute GVHD, 2 developed degree Ⅰ , one developed degree Ⅱ , and one developed degree Ⅳ. Seven patients were alive 100 days after allo-HSCT, 3 of whom presented chronic GVHD. At the end of following-up period, 6 patients were alive, among them, 3 patients were alive without relapse; 3 patients relapsed; Two patients died, one from acute GVHD, and one from leukemia relapse. Conclusion Combined intensive preconditioning regimen allo-HSCT with Imatinib was an effective treatment for Ph+ ALL, but the effect of anti-chronic GVHD of imatinib should arouse certain attention.

Objective To observe the effects of bone marrow?derived mesenchymal stem cells (BMSC) on glomerular podocyte injured by lipopolysaccharide (LPS) and the expression of related protein. Methods Podocytes are divided into control group, BMSC group, LPS group and LPS plus BMSC group. After 24 hours of intervention, observing each experimental group podocyte form under inverted phase contrast microscope;detecting the expressions of mRNA and protein of nephrin, CD2AP, synaptopodin, and TRPC6 by RT?PCR and Western?blot. Results Compared with control group, expressions of nephrin, CD2AP, and synaptopodin in LPS group decreased (P<0.05) while that of TRPC6 increased (P<0.05); compared with LPS group, expressions of nephrin, CD2AP, and synaptopodin in LPS+MSC group increased (P<0.05) while that of TRPC6 decreased (P<0.05). Conclusion BMSC may relieve LPS?induced podocyte injury.

Lipase gene from Penicillium expansum(lip07) was cloned and over-expressed in Pichia pastoris.a random mutant named ep8,which contained a single amino acid substitution,was obtained by using the lip07 as an error-prone PCR template in previous study.ep8 shows higher thermostability than that of lip07,To further improve the thermostability of the lipase,the Lys of wild-type(lip07) and mutant(ep8) in 202 were substituted by Ala using the Overlap extension PCR technique respectively.The mutant genes(lip07-K202A and ep8-K202A) were subcloned into pAO815,and then transformed into the Pichia pastoris GS115 for extracelluar expression,respectively.15% SDS-PAGE analysis indicated that the molecular mass of PEL-ep8-K202A and PEL-lip07-K202A are both about 28kDa,which is same with the wild-type lipase.The Tm of PEL-ep8-K202A is 41.66℃,2.63℃ higher than that of the wild-type(39.03℃) and 1.21℃ higher than the random mutant(PEL-ep8:40.45℃);the Tm of single mutant(PEL-lip07-K202A) is 37.08℃,2℃ lower than that of the wild-type lipase.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Neoplasm Invasiveness ; Neoplasm Metastasis ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

OBJECTIVE: To construct the plasmid of human vascular endothelial cell growth factor165 and green fluorescence protein report gene eukaryotic expression vector of fusion protein pEGFP /hVEGF165, and to detect its expression in vascular endothelial cells. METHODS: We amplified full-length of gene VEGF165 by PCR, cloned in direction in multiple clone sites of pEGFP-N1, constructed recombinant plasmid of pEGFP/hVEGF165. Through enzyme digestion, PCR, and sequencing analysis, we also performed liposome-mediated transfection of vascular endothelial cells of in vitro cultivation, and detected the expression of fusion protein pEGFP/hVEGF165 using fluorescence microscope, RT-PCR, and Western blot. RESULTS: Both gene VEGF165 and multiple clone site of pEGFP-N1 confirmed by PCR, enzyme digestion, and sequence analysis. EGFP/VEGF protein was expressed in vascular endothelial cells after pEGFP/VEGF165 recombinant plasmid transfected vascular endothelial cells. CONCLUSION Fusion protein eukaryotic plasmid of report gene EGFP and VEGF165 is successfully constructed, and EGFP/VEGF can be expressed in vascular endothelial cells, which lays a foundation for the application of VEGF gene in treating ischemia vascular diseases.
Cells, Cultured ; Endothelial Cells ; metabolism ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Plasmids ; RNA, Messenger ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo observe the effects of Danshen root compound (DSC) on blood lipid and bone biomechanics in mice with hyperlipemia-induced osteoporosis.

METHODSForty Kunming mice were randomized into 5 equal groups, and were given intragastric administration with distilled water (control), lipid emulsion (LE) at the daily dose of 5 ml/kg, LE plus simvastatin, LE plus DSC at 5.0 g/kg (DSC-L group), and LE plus DSC at 10.0 g/kg (DSC-H group), respectively. Serum TC, TG, and HDL-c levels and left femur hydroxyproline, calcium and phosphate contents were measured in the rats, with the right femur taken for bone biomechanical test.

RESULTSCompared with those in the control group, serum TC, LDL-c and AI of the mice increased and HDL-c, Hyp and bone calcium decreased significantly (P<0.01) with lowered bone biomechanical properties. Compared with those of the LE model group, AI decreased and HDL-c increased significantly in DSC-L and DSC-H groups (P<0.01), and the bone biomechanics in DSC-H group was improved.

CONCLUSIONLong-term intragastric administration of lipid emulsion causes lipid metabolic disorder and induces osteoporosis due to hyperlipemia in mice. DSC can significantly increase HDL-c and partially prevent the occurrence of osteoporosis in mice.

Animals ; Biomechanical Phenomena ; Bone and Bones ; drug effects ; metabolism ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Hyperlipidemias ; complications ; drug therapy ; Lipids ; blood ; Male ; Mice ; Osteoporosis ; blood ; etiology ; prevention & control ; Phenanthrolines ; pharmacology ; therapeutic use ; Phytotherapy ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo investigate the preventive effects of a compound Danshen preparation (DSC) on long-term gastric lipid emulsion administration-induced nonalcoholic steatohepatitis in rats.

METHODSTwenty-seven 3-month-old SD rats were randomized equally into 3 groups and subjected to daily intragastric administration for 20 weeks of distilled water (control), lipid emulsion at 5 ml/kg (model group), and lipid emulsion plus DSC at 5.0 g/kg (DSC treatment group). After blood glucose (BG) determination, the rats were sacrificed for measurement of serum TC, TG, HDL-c, AST, and ALT, and the liver was weighed and pathologically examined.

RESULTSCompared with the control group, the rats in the model group showed significantly increased BG, TC, LDL-c, arteriosclerosis index (AI), AST, ALT, liver weight, and liver index (P<0.01) and decreased HDL-c (P<0.01), while TG remained unchanged. Fatty degeneration, hydropic degeneration and necrosis with inflammatory cell infiltration were observed in the liver of the rats in the model group. Compared with the model group, the rats in DSC groups showed decreased BG, AI (P<0.01), liver weight, liver index, AST, and ALT (P<0.05) and increased HDL-c, with milder pathological changes in the liver.

CONCLUSIONSLong-term gastric perfusion of lipid emulsion causes lipid metabolic disorder and nonalcoholic fatty liver disease in rats characterized by increased TC and decreased HDL-c. DSC can significantly increase HDL-c and provide partial protection of the liver against the damages by the lipid emulsion.

Animals ; Drug Administration Routes ; Drugs, Chinese Herbal ; therapeutic use ; Emulsions ; Fatty Liver ; chemically induced ; prevention & control ; Female ; Lipids ; administration & dosage ; toxicity ; Male ; Phenanthrolines ; therapeutic use ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; Time Factors